Frans_M_Everaerts_Isotachophoresis_378342.pdf

360 ENZYMATIC REACTIONS
TABLE 15.9
MASS BALANCE OF: THE REACTION CONSTITUENTS
Reaction time Amount (nmole)
(min)
-~___
Pyruvate Lactate NADH NAD
40 - 148.4 134.8 - 127.2 128.2
60 - 195.5 196.4 - 180.6 183.4
70 -206.3 227.2 - 197.8 205.5
80 -235.3 250.3 -218.2 225.4
method (Chapter 10) because only two componentsneed to be quantitatively determined.
The results of these experiments are given in Table 15.9.
Table 15.9 shows that an acceptable agreement is obtained, although it is not clear
why the values for pyruvate and lactate on the one hand and for NADH and NAD on
the other are so similar. The reason may lie in impurities in the standards used for the
calibration, and especially impurities present in NAD and NADH. All enzymatic reactions
in which NAD or NADH are involved can be analyzed by isotachophoresis [7].
REFERENCES
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2 H.J. Fromm and V. Zewe,J. Biol. Chern., 237 (1962) 3027.
3 H.J. Frornm, J. Biol. Chem., 239 (1964) 3045.
4 P. Ottolenghi, C.R. Trav. Lab. Carlsberg, 34 (1964) 237.
5 N.C. Melchior and J.B. Melchior, J. Biol. Chern., 231 (1958) 609.
6 J. Hoenkamp, Graduation Rep., University of Technology, Eindhoven, 1975.
7 T. Willemsen, Graduation Rep., University of Technology, Eindhoven, 1974.